Project description:Mice of indicated genotypes were perfused and their brains dissected and dissociated. Cells were fixed, immunolabeled and FACS sorted. RNA was extracted from neuron, astrocyte, and microglial cell populations. Typical RIN=4-5 for neurons, 6-8 for astrocytes, and 5-7 for microglia. Typical RNA yields 200-300ng for neurons, ~20ng for microglia, and ~5ng for astrocytes. cDNA was generated from up to 25 ng of total RNA using Nugen’s RNA-Seq method for low-input RNA samples, Ovation RNA-Seq System V2 (NuGEN). (Per manufacturers instructions, total RNA was neither depleted of rRNA nor polyA-selected.) 1 μg of sheared cDNA was taken into further processing, starting at end repair step, using Illumina’s TruSeq RNA Sample Preparation Kit v2 (Illumina). The SAMPLE_ID sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0006717
Project description:Comparing Trem2-KO;PS2APP and Trem2-WT;PS2APP CD11b+ cells reveals the role of Trem2 in microglial gene expression in amyloid-laden brains. The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0014430
Project description:RNA was purified from intact cerebrocortical tissue of female PS2APP or non-transgenic mice, perfused at 7 or 13 months of age. The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0007648
Project description:This project defines the microglial gene expression profile in a transgenic mouse model of Alzheimer's, compared to non-transgenic age-matched controls, at a time when amyloid pathology and microgliosis are rampant. Microglia were sorted live from one hemisphere of cerebral cortex, using GFP expressed from Cx3cr1 locus (mice have one intact copy of Cx3cr1). RNA was isolated from sorted microglia using RNeasy mini. Two groups, PS2APP and non-transgenic, with 9 mice/group, aged 14-15 months. The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0006715
Project description:Here we studied cellular level changes of hippocampal cells by unbiased single cell RNA-seq using AD mouse models bearing amyloid pathology (PS2APP), or amyloid and tau combined pathology (TauPS2APP) by single cell RNA-seq. We identified 16 different cell types and further characterized responses of microglia, oligodendrocytes, astrocytes and T cells to amyloidosis only and amyloid plus tau combined pathology. Both mouse models exhibited robust microglial responses. We also found distinct responses of oligodendrocytes to different AD pathologies. We observed increased T cell numbers in both mouse models. Current dataset presents diverse transcriptomic responses to AD pathology and provides resources to study molecular mechanisms underlying disease onset and progression.
Project description:Affymetrix miRNA arrays were used to generate miRNA profiles of peripheral blood leukocytes and FACS sorted neutrophils, monocytes, B-cells, T-cells, CD4+ T-cells, and CD8+ T-cells, being the major leukocyte cell types in human. The study allowed for the determination of the miRNAs that were expressed in each leukocyte cell subtype. Two-way hierarchical clustering on the miRNAs and samples illustrated that miRNA expression profiles of B- and T-cells were very much alike, and that there there were a number of miRNAs which appeared to have an expression profile specific to certain leukocyte cell subtypes. This study will facilitate the identification of microRNAs associated with and contributing to single leukocyte cell subtypes.
Project description:Affymetrix miRNA arrays were used to generate miRNA profiles of peripheral blood leukocytes and FACS sorted neutrophils, monocytes, B-cells, T-cells, CD4+ T-cells, and CD8+ T-cells, being the major leukocyte cell types in human. The study allowed for the determination of the miRNAs that were expressed in each leukocyte cell subtype. Two-way hierarchical clustering on the miRNAs and samples illustrated that miRNA expression profiles of B- and T-cells were very much alike, and that there there were a number of miRNAs which appeared to have an expression profile specific to certain leukocyte cell subtypes. This study will facilitate the identification of microRNAs associated with and contributing to single leukocyte cell subtypes. Peripheral blood leukocytes were extracted, and neutrophils, monocytes, B-cells, T-cells, CD4+ T-cells, and CD8+ T-cells were FACS sorted from total blood from a healthy subject. In order to determine the miRNA expression profile of these cells, RNA was extracted, labeled and hybridized on an Affymetrix miRNA array.
Project description:Mice of indicated ages and genotypes were perfused and their brains dissected and dissociated. Cells were fixed, immunolabeled and FACS sorted. RNA was extracted from neuron, astrocyte, and microglial cell populations. Typical RIN=4-5 for neurons, 6-8 for astrocytes, and 5-7 for microglia. Typical RNA yields ~100ng for neurons, ~20ng for microglia, and ~10ng for astrocytes. cDNA was generated from up to 25 ng of total RNA using Nugen's RNA-Seq method for low-input RNA samples, Ovation RNA-Seq System V2 (NuGEN). (Per manufacturers instructions, total RNA was neither depleted of rRNA nor polyA-selected.) 1 ug of sheared cDNA was taken into further processing, starting at end repair step, using Illumina's TruSeq RNA Sample Preparation Kit v2 (Illumina). The 'SAMPLE_ID'; sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0006149 Astrocytes, microglia and neurons were sorted from 7- or 13-month old PS2APP or non-transgenic mice, 4 <= n <= 7 per group.